Observations of Cell Size Dynamics Under Osmotic Stress
Observations of Cell Size Dynamics Under Osmotic Stress
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DOI：
10.1002/cyto.a.21076
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发表时间：
2011-07-01
期刊：
影响因子：
3.7
通讯作者：
Sharfstein, Susan T.
中科院分区：
文献类型：
--
作者：
Kiehl, Thomas R.;Shen, Duan;Sharfstein, Susan T.
用于产生治疗和诊断蛋白的培养哺乳动物细胞[如小鼠杂交瘤、中国仓鼠卵巢(CHO)细胞]在渗透胁迫下往往表现出更高的比产。这种比生产力的增加伴随着许多生理变化，包括细胞大小的变化。研究高渗胁迫下培养物细胞大小的变化，可能在一定程度上揭示提高特定生产力的基础，以及了解在高渗条件下发生的一些细胞防御反应。细胞体积的调节是维持在动物细胞中的一项重要功能。尽管这些电池对水具有很高的渗透性，但它们对离子溶质的渗透性却明显较低。在这些细胞中，通过调节离子和渗透分子的平衡来积极地维持适当的细胞水分含量。适合细胞外条件的运输，导致这些物种的积累或释放，是对急性细胞体积变化的反应。渗透压诱导的调节性容量增加(RVI)和调节性容量减少(RVD)是已知在各种条件下发生的。我们观察了两种CHO细胞系在高渗条件下群体大小变化的时间演化。使用多种仪器、多种细胞系和多种细胞培养条件进行观察。使用LSRII(R)流式细胞仪通过流式细胞仪测量CHO A1的大小变化，而使用Cedex(R)细胞分析仪对CHO B0细胞进行定量。高渗应激对细胞体积的调节作用具有剂量依赖性。悬浮培养的CHO细胞的平均细胞直径发生了变化。这种平均值的变化不是由于整个种群的变化，而是因为出现了细胞亚群的不同亚群，其细胞直径比大多数种群中的细胞直径大。(C)2011年国际细胞测量促进会
Cultured mammalian cells [e.g., murine hybridomas, Chinese hamster ovary (CHO) cells] used to produce therapeutic and diagnostic proteins often exhibit increased specific productivity under osmotic stress. This increase in specific productivity is accompanied by a number of physiological changes, including cell size variation. Investigating the cell size variation of hyperosmotically stressed cultures may reveal, in part, the basis for increased specific productivity as well as an understanding of some of the cellular defense responses that occur under hyperosmotic conditions. The regulation of cell volume is a critical function maintained in animal cells. Although these cells are highly permeable to water, they are significantly less permeable to ionic solutes. Appropriate cell water content is actively maintained in these cells by regulation of ion and osmolyte balances. Transport appropriate to extracellular conditions, leading to accrual or release of these species, is activated in response to acute cell volume changes. Osmotically induced regulatory volume increases (RVI) and regulatory volume decreases (RVD) are known to occur under a variety of conditions. We observed the time evolution of size variation in populations of two CHO cell lines under hyperosmotic conditions. Observations were made using multiple instruments, multiple cell lines, and multiple cell culture conditions. Size variation of CHO A1 was gauged by flow cytometry using an LSRII (R) flow cytometer while CHO B0 cells were quantified using a Cedex (R) cell analyzer. Hyperosmotic stress had a dose-dependent effect on the regulatory control of cell volume. Stressed cultures of CHO cells grown in suspension exhibited a shift in mean cell diameter. This shift in mean was not due to a change in the whole population, but rather to the emergence of distinct subpopulations of cells with larger cell diameters than those in the bulk of the population. (C) 2011 International Society for Advancement of Cytometry